LSD1 inhibitors and medical uses thereof

ABSTRACT

Provided are novel compounds of Formula (I or Ia′): and pharmaceutically acceptable salts thereof, which are useful for treating a variety of diseases, disorders or conditions, associated with LSD1. Also provided are pharmaceutical compositions comprising the novel compounds of Formula (I or Ia′), pharmaceutically acceptable salts thereof, and methods for their use in treating one or more diseases, disorders or conditions, associated with LSD1.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/345,011, filed Apr. 25, 2019, which is a 35 U.S.C. § 371 nationalstage filing International Application No. PCT/US2017/058405, filed Oct.26, 2017, which claims priority to U.S. Provisional Application No.62/413,162, filed Oct. 26, 2016 and U.S. Provisional Application No.62/413,166, filed Oct. 26, 2016, the contents of each of which areincorporated herein by reference.

BACKGROUND

Lysine-specific demethylase (LSD1), also known as lysine (K)-specificdemethylase 1A (LSD1), is a protein in humans that in encoded by theKDM1A gene and specifically demethylates mono- or dimethylated histoneH3 lysine4 (H3K4) and H3 lysine 9 (H3K9) via a redox process. Biochimicaet Biophysica Acta 1829 (2013) 981-986. LSD1 has been found to possessoncogenic properties in several cancers ranging from prostate (CancerRes., 66 (2006), pp. 11341-11347) bladder (Mol. Carcinog., 50 (2011),pp. 931-944) neuroblastomas, (Cancer Res., 69 (2009), pp. 2065-2071)lung cancers, (PLoS One, 7 (2012), p. e35065) sarcomas andhepato-carcinomas (Tumour Biol. (2012). LSD1 pharmacological inhibitorshave been shown e.g., to treat leukemias (Nat. Med., 18 (2012), pp.605-611) and also solid tumors (Tumour Biol. (2012)).

SUMMARY

It has now been found that compounds of structural Formula:

or a pharmaceutically acceptable salt thereof, and compositionscomprising compounds of this Formula, wherein each of R¹, R², R³, R⁴,R⁵, R⁶, m, n, o, and q are defined and described herein, are effectiveinhibitors of LSD1. See Table 6 in the Exemplification Section below.Conditions treated by the disclosed compounds are described herein.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts the X-ray crystal structure for intermediate(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate.

FIG. 2 depicts the X-ray crystal structure for the monocitrate salt ofCompound 2.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. General Description ofCompounds

In certain embodiments, the present disclosure provides a compound ofFormula I:

or a pharmaceutically acceptable salt thereof, wherein

n is 1 or 2;

m is 1 or 2;

o is 0 or 1;

q is 1, 2, or 3;

R¹ is hydrogen or (C₁-C₆)alkyl;

R² is hydrogen or (C₁-C₆)alkyl optionally substituted with OH or(C₁-C₆)alkoxy;

R³ and R⁴, if present are each independently selected from hydrogen,halo, OH, and (C₁-C₆)alkyl;

R⁵ is selected from NH₂, —NH(SO₂)(C₁-C₆)alkyl,—NH(SO₂)(C₁-C₆)alkylO(C₁-C₆)alkyl, —NHC(O)(C₁-C₆)alkyl,—NH(SO₂)(C₃-C₆)cycloalkyl, OH, —O(C₁-C₆)alkyl, —SO₂NH₂, —C(O)NH₂,—C(O)NH(C₁-C₆)alkyl, —C(O)N[(C₁-C₆)alkyl]₂, —C(O)NH(SO₂)(C₁-C₆)alkyl,—C(O)NH(C₁-C₆)alkyl(SO₂)(C₁-C₆)alkyl, —C(O)NH(SO₂)(C₃-C₆)cycloalkyl,—C(O)NH(C₁-C₆)alkylOH, —C(O)NH(C₁-C₆)alkylO(C₁-C₆)alkyl, and(C₁-C₄)alkyl substituted with OH; and

R⁶ is selected from (C₁-C₆)alkyl, halo(C₁-C₆)alkyl, (C₁-C₆)alkoxy,halo(C₁-C₆)alkoxy, halo, and cyano.

In certain embodiments, the present disclosure provides a compound ofFormula Ia′:

or a pharmaceutically acceptable salt thereof, wherein

n is 1 or 2;

m is 1 or 2;

o is 0 or 1;

q is 0, 1, 2, or 3;

R¹ is hydrogen or (C₁-C₆)alkyl;

R² is hydrogen or (C₁-C₆)alkyl optionally substituted with OH or(C₁-C₆)alkoxy;

R³ and R⁴, if present are each independently selected from hydrogen,halo, OH, and (C₁-C₆)alkyl;

R⁵ is selected from NH₂, —NH(SO₂)(C₁-C₆)alkyl,—NH(SO₂)(C₁-C₆)alkylO(C₁-C₆)alkyl, —NHC(O)(C₁-C₆)alkyl,—NH(SO₂)(C₃-C₆)cycloalkyl, OH, —O(C₁-C₆)alkyl, —SO₂NH₂, —C(O)NH₂,—C(O)NH(C₁-C₆)alkyl, —C(O)N[(C₁-C₆)alkyl]₂, —C(O)NH(SO₂)(C₁-C₆)alkyl,—C(O)NH(C₁-C₆)alkyl(SO₂)(C₁-C₆)alkyl, —C(O)NH(SO₂)(C₃-C₆)cycloalkyl,—C(O)NH(C₁-C₆)alkylOH, —C(O)NH(C₁-C₆)alkylO(C₁-C₆)alkyl, and(C₁-C₄)alkyl substituted with OH; and

R⁶, if present, is selected from (C₁-C₆)alkyl, halo(C₁-C₆)alkyl,(C₁-C₆)alkoxy, halo(C₁-C₆)alkoxy, halo, and cyano.

2. Compounds and Definitions

The terms “halo” and “halogen” as used herein refer to an atom selectedfrom fluorine (fluoro, F), chlorine (chloro-, Cl), bromine (-bromo, Br),and iodine (iodo-, -I).

The term “alkyl” used alone or as part of a larger moiety, such as“alkoxy”, “haloalkyl”, and the like, means saturated straight-chain orbranched monovalent hydrocarbon radical. Unless otherwise specified, analkyl group typically has 1-6 carbon atoms, i.e., (C₁-C₆)alkyl. As usedherein, a “(C₁-C₆)alkyl” group is means a radical having from 1 to 6carbon atoms in a linear or branched arrangement.

The term “haloalkyl” includes mono, poly, and perhaloalkyl groups wherethe halogens are independently selected from fluorine, chlorine,bromine, and iodine.

“Alkoxy” means an alkyl radical attached through an oxygen linking atom,represented by —O-alkyl. For example, “(C₁-C₄)alkoxy” includes methoxy,ethoxy, proproxy, and butoxy.

When used in connection to describe a chemical group that may havemultiple points of attachment, a hyphen (-) designates the point ofattachment of that group to the variable to which is is defined. Forexample, —NH(SO₂)(C₃-C₆)cycloalkyl means that the point of attachmentfor this group is on the nitrogen atom.

The disclosed compounds exist in various stereoisomeric forms.Stereoisomers are compounds that differ only in the spatial arrangementof their atoms. Different spatial arrangements in a compound can resultfrom e.g., the orientation of four different substituents around achiral carbon atom (i.e., a chiral center), the orientation of two ormore substituents around a double bond, or the orientation of two ormore substituents on a cycloalkyl ring.

Enantiomers are one type of stereoisomer that can arise from a chiralcenter or chiral centers. Enantiomers are pairs of stereoisomers whosemirror images are not superimposable, most commonly because they containan asymmetrically substituted carbon atom or carbon atoms that acts as achiral center(s). “R” and “S” represent the absolute configuration ofsubstituents around one or more chiral carbon atoms, where each chiralcenter is assigned the prefix “R” or “S” according to whether the chiralcenter configuration is right- (clockwise rotation) or left-handed(counter clockwise rotation). If the turn is clockwise or right-handedabout a chiral carbon, the designation is “R” for rectus. If the turn iscounter clockwise or left-handed about a chiral carbon, the designationis “S” for sinister.

Enantiomeric purity reflects the degree to which one enantiomer of acompound is predominantly present over the other enantiomer of thatcompound. It is determined by subtracting the percent composition of themajor enantiomer with the percent composition of the minor enantiomerthat is present. For example, a racemic mixture has an enantiomericpurity of 0%, while a single completely pure enantiomer has anenantiomeric purity of 100%. A composition with 70% of one enantiomerand 30% of the other has an enantiomeric purity of 40% (70%-30%).

Diastereomers are stereoisomers that are not related as object andmirror image and are not enantiomers. Unlike enantiomers which aremirror images of each other and non-superimposable, diastereomers arenot mirror images of each other and non-superimposable. Diastereomershave two or more chiral centers.

Geometric isomers arise when two or more substituents on a double bondor ring can have different spatial orientations with respect to oneanother due to the presence of the double bond or ring structure. Whenthe orientation of the substituents of a geometric isomer are onopposite sides of the double bond, those substituents are said to be“trans” to one another or denoted by the letter “E.” When theorientation of the substituents of a geometric isomer are on the sameside of the double bond, those substituents are said to be “cis” to oneanother or denoted by the letter “Z.”

When the configuration of two or more substituents about a double bondis indicated by structure; by “E” or “Z” designations; by “cis” or“trans”; or by a combination of the foregoing, it is to be understoodthat the depicted stereochemical purity with respect to that double bondis at least 85%, at least 90%, at least 95%, at least 97%, at least 98%or at least 99% by weight. Stereochemical purity by weight with respectto a double bond means the percent by weight of the compound in acomposition having the indicated stereochemistry about the double bond.For example, in compounds having the Formula I or Ia′, when the doublebond is represented by

it is to be understood that the compound has a stereochemical puritywith respect to the depicted trans (E) stereochemistry about the doublebond, i.e., at least 85%, at least 90%, at least 95%, at least 97%, atleast 98% or at least 99% by weight of the compound in a compositioncontains the represented trans (i.e., E) double bond.

Geometric isomers can also arise based on the orientation of two or moresubstituents about a cyclic group. For example, in compounds of FormulaI or Ia′, the orientation of

about the cyclopropyl can give rise to two different cis configurations(as in

and two different trans configurations (as in

In instances where the stereochemistry about the cyclopropyl is notdefined, as in

the structure includes one of the cis or trans isomers free of other cisand trans stereoisomers, or, alternatively, any mixture of cis and transstereoisomers.

When the stereochemistry about the cyclopropyl in the compounds ofFormula I or Ia′ is indicated by structure only, the structure is meantto depict the relative stereochemistry at one of the chiral centers inthe cyclopropyl relative to the stereochemistry at the other chiralcenter, and not the absolute stereochemistry at either chiral center inthe cyclopropyl. For example, when the stereochemistry about thecyclopropyl is depicted by structure only as being trans, thestereochemical purity of the compound with respect to the depicted transconfiguration about the cyclopropyl is at least 85%, at least 90%, atleast 95%, at least 97%, at least 98% or at least 99% by weight, i.e.,the percent by weight of the compound in the composition having thetrans stereochemistry at the cyclopropyl is at least 85%, at least 90%,at least 95%, at least 97%, at least 98% or at least 99% by weight. Forexample, a compound represented by the formula:

means that at least 85%, at least 90%, at least 95%, at least 97%, atleast 98% or at least 99% by weight of the compound in the compositionhas the depicted trans configuration about the cyclopropyl; at least atleast 85%, at least 90%, at least 95%, at least 97%, at least 98% or atleast 99% by weight of the compound in the composition contains theother trans configuration as:

or at least 85%, at least 90%, at least 95%, at least 97%, at least 98%or at least 99% by weight of the compound in the composition is amixture of the two trans configurations.

When the absolute stereochemistry of chiral centers in a compound areindicated structurally and by “R” or “S” designations, it is to beunderstood that the depiction means the depicted stereoisomer at astereochemical purity of at least 85%, at least 90%, at least 95%, atleast 97%, at least 98% or at least 99% by weight, i.e., the percent byweight of the indicated stereoisomer of the compound in the composition.For example, a compound of Formula I or Ia′ represented by the formula:

means at least 85%, at least 90%, at least 95%, at least 97%, at least98% or at least 99% by weight of the compound of Formula I in thecomposition contains of the depicted stereoisomer. When the structurebeing depicted by structure and by “R” or “S” designation is a singleenantiomer, the enantiomeric purity is at least 95% (e.g., at least 96%,at least 97%, at least 98%, at least 99%, at least 99.5%, or at least99.9%).

When a compound is depicted by structurally without indicating thestereochemistry at a chiral center, it is to be understood that thestructure includes either configuration at the chiral center or,alternatively, any mixture of configurations at that chiral center.

The 1- and 2-positions of the cyclopropyl ring represent the following:

The compounds described herein may be present in the form ofpharmaceutically acceptable salts. For use in medicines, the salts ofthe compounds of the invention refer to non-toxic “pharmaceuticallyacceptable salts.” Pharmaceutically acceptable salt forms includepharmaceutically acceptable acidic/anionic or basic/cationic salts.Suitable pharmaceutically acceptable acid addition salts of thecompounds described herein include e.g., salts of inorganic acids (suchas hydrochloric acid, hydrobromic, phosphoric, nitric, and sulfuricacids) and of organic acids (such as, acetic acid, benzenesulfonic,benzoic, methanesulfonic, and p-toluenesulfonic acids). Compounds of thepresent teachings with acidic groups such as carboxylic acids can formpharmaceutically acceptable salts with pharmaceutically acceptablebase(s). Suitable pharmaceutically acceptable basic salts include e.g.,ammonium salts, alkali metal salts (such as sodium and potassium salts)and alkaline earth metal salts (such as magnesium and calcium salts).Compounds with a quaternary ammonium group also contain a counteranionsuch as chloride, bromide, iodide, acetate, perchlorate and the like.Other examples of such salts include hydrochlorides, hydrobromides,sulfates, methanesulfonates, nitrates, benzoates and salts with aminoacids such as glutamic acid.

The terms “subject” and “patient” may be used interchangeably, and meansa mammal in need of treatment, e.g., companion animals (e.g., dogs,cats, and the like), farm animals (e.g., cows, pigs, horses, sheep,goats and the like) and laboratory animals (e.g., rats, mice, guineapigs and the like). Typically, the subject is a human in need oftreatment.

3. Description of Exemplary Compounds

In a first embodiment, the present disclosure provides a compound ofFormula I or Ia′:

or a pharmaceutically acceptable salt thereof, wherein the variables areas described above.

In a second embodiment, the present disclosure provides a compound ofFormula Ia:

or a pharmaceutically acceptable salt thereof, wherein the variables areas described above.

In a third embodiment, R⁶ in Formula I, Ia′, or Ia is selected frommethyl, ethyl, halomethyl, haloethyl, methoxy, halomethoxy, halo, andcyano, wherein the remaining variables are as described in Formula I,Ia′, or the second embodiment.

In a fourth embodiment, the compound of Formula I, Ia′, or Ia is of theFormula II:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a fifth embodiment, the compound of Formula I, Ia′, Ia, or II is ofthe Formula III:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a sixth embodiment, the compound of Formula I, Ia′, Ia, or II is ofthe Formula IV:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a seventh embodiment, the compound of Formula I, Ia′, Ia, II, or IIIis of the Formula V:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In an eighth embodiment, R² in the compounds of Formula I, Ia′, Ia, II,III, IV, or V is hydrogen, methyl, or hydroxy(C₁-C₄)alkyl, wherein theremaining variables are as described in Formula I or in the thirdembodiment. Alternatively, R² in the compound of Formula I, Ia′, Ia, II,III, IV, or V is hydroxy(C₁-C₄)alkyl, wherein the remaining variablesare as described in Formula I and the third embodiment. In anotheralternative, R² in the compound of Formula I, Ia′, Ia, II, III, IV, or Vis hydroxy(C₁-C₂)alkyl, wherein the remaining variables are as describedin Formula I and the third embodiment.

In a ninth embodiment, the compound of Formula I, Ia′, Ia, II, or III isof the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a tenth embodiment, the compound of Formula I, Ia′, Ia, II, or III isof the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In an eleventh embodiment, the compound of Formula I, Ia′, Ia, II, orIII is of the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a twelfth embodiment, the compound of Formula I, Ia′, Ia, II, or IVis of the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a thirteenth embodiment, the compound of Formula I, Ia′, Ia, II, IVor IX is of the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a fourteenth embodiment, the compound of Formula I, Ia′, Ia, II, IV,or IX is of the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a fifteenth embodiment, the compound of Formula I, Ia′, Ia, II, IV,or IX is of the Formula:

or a pharmaceutically acceptable salt thereof, wherein the remainingvariables are as described in Formula I or Ia′.

In a sixteenth embodiment, R⁵ in any one of the compounds of Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII is NH₂,—NH(SO₂)(C₁-C₄)alkyl, —NH(SO₂)(C₁-C₄)alkylO(C₁-C₄)alkyl,—NHC(O)(C₁-C₄)alkyl, —NH(SO₂)(C₃-C₆)cycloalkyl, OH, —SO₂NH₂, —C(O)NH₂,—C(O)NH(C₁-C₄)alkyl, —C(O)NH(SO₂)(C₁-C₄)alkyl,—C(O)NH(C₁-C₄)alkyl(SO₂)(C₁-C₄)alkyl, —C(O)NH(C₁-C₄)alkylOH, or(C₁-C₂)alkyl substituted with OH, wherein the remaining variables are asdescribed for Formula I or Ia′ and the third or eighth embodiments.Alternatively, R⁵ in any one of the compounds of Formula I, Ia′, Ia, II,III, IV, V, VI, VII, VIII, IX, X, XI, or XII is NH₂, —NH(SO₂)CH₃,—NH(SO₂)(CH₂)₂OCH₃, —NH(SO₂)CH(CH₃)₂, —NH(SO₂)CH₂CH₃, —NHC(O)CH₃,—NH(SO₂)cyclopropyl, OH, —SO₂NH₂, —C(O)NH₂, —C(O)NH(SO₂)CH₃,—C(O)NH(CH₃)₂, —C(O)NHCH(CH₃)₂, —C(O) NHCH₂CH₃, —C(O)NH(CH₂)₂(SO₂)CH₃,—C(O)NH(CH₂)₂OH, or —CH₂OH, wherein the remaining variables are asdescribed for Formula I or Ia′, and the third or eighth embodiments. Inanother alternative, R⁵ in any one of the compounds of Formula I, Ia′,Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII is —C(O)NH₂, OH,—SO₂NH₂, or —NH(SO₂)CH₃, wherein the remaining variables are asdescribed for Formula I or Ia′ and the third or eighth embodiments. Inyet another alternative, R⁵ in any one of the compounds of Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII is OH, whereinthe remaining variables are as described for Formula I or Ia′ and thethird or eighth embodiments.

In a seventeenth embodiment, the compound of Formula Ia′ is selectedfrom from

or a pharmaceutically acceptable salt thereof.

In an eighteenth embodiment, the compound of Formula Ia′ is:

or a pharmaceutically acceptable salt thereof.

In a nineteenth embodiment, the compound of Formula Ia′ is:

or a pharmaceutically acceptable salt thereof.

In a twentieth embodiment, the compounds described herein (e.g., FormulaI, Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII and thosein the seventeenth to nineteenth embodiment) are single enantiomershaving an enantiomeric purity of at least 95% (e.g., at least 96%, atleast 97%, at least 98%, at least 99%, at least 99.5%, or at least99.9%).

In a twenty-first embodiment, the stereochemical configuration about thecyclopropyl ring in the compounds described herein (e.g., Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII and those inthe seventeenth to nineteenth embodiment) is 1R,2S.

In a twenty-second embodiment, the stereochemical configuration aboutthe cyclopropyl ring in the compounds described herein (e.g., Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII and those inthe seventeenth to nineteenth embodiment) is 1R,2S and the compounds aresingle enantiomers having an enantiomeric purity of at least 95% (e.g.,at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%,or at least 99.9%).

In a twenty-third embodiment, the stereochemical configuration about thecyclopropyl ring in the compounds described herein (e.g., Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII and those inthe seventeenth to nineteenth embodiment) is 1S,2R.

In a twenty-fourth embodiment, the stereochemical configuration aboutthe cyclopropyl ring in the compounds described herein (e.g., Formula I,Ia′, Ia, II, III, IV, V, VI, VII, VIII, IX, X, XI, or XII and those inthe seventeenth to nineteenth embodiment) is 1S,2R and the compounds aresingle enantiomers having an enantiomeric purity of at least 95% (e.g.,at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%,or at least 99.9%).

In one aspect, the compounds described herein are not

or a salt thereof.

In one aspect, when q is 0, m is 1, n is 1, o is 0, and R¹ and R² areboth hydrogen in the compounds described herein then R⁵ is not —CONH₂.In another aspect, when q is 0, m is 2, n is 2, o is 0, and R¹ and R²are both hydrogen in the compounds described herein then R⁵ is not—CONH₂.

Specific examples of compounds are provided in the EXEMPLIFICATIONsection and are included as part of a fourteenth embodiment herein.Pharmaceutically acceptable salts as well as the neutral forms of thecompounds in the EXEMPLIFICATION are also included.

4. Formulation and Administration

In one aspect, provided herein are compositions comprising the compoundsdescribed herein, or a pharmaceutically acceptable salt thereof.

In another aspect, the compositions further comprise a pharmaceuticallyacceptable carrier, adjuvant, or vehicle. The amount of compound in thecomposition is such that is effective to measurably modulate LSD1, or amutant thereof in a biological sample or in a patient, i.e., the“effective amount” or “therapeutically effective amount.”

In certain aspects, a composition described herein is formulated foradministration to a patient in need of such composition. In someaspects, a composition described herein is formulated for oraladministration to a patient.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle” or““pharmaceutically acceptable carrier” refers to a non-toxic carrier,adjuvant, or vehicle that does not destroy the pharmacological activityof the compound with which it is formulated. Pharmaceutically acceptablecarriers, adjuvants or vehicles that may be used in the compositionsdescribed herein include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins, such as humanserum albumin, buffer substances such as phosphates, glycine, sorbicacid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

Pharmaceutically acceptable compositions described herein may he orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Pharmaceutically acceptable compositions described herein may also beprepared in injectable form. Injectable preparations, for example,sterile injectable aqueous or oleaginous suspensions may be formulatedaccording to the known art using suitable dispersing or wetting agentsand suspending agents. The sterile injectable preparation may also be asterile injectable solution, suspension or emulsion in a nontoxicparenterally acceptable diluent or solvent, for example, as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution, U.S.P. and isotonic sodiumchloride solution. In addition, sterile, fixed oils are conventionallyemployed as a solvent or suspending medium. For this purpose any blandfixed oil can be employed including synthetic mono- or diglycerides. Inaddition, fatty acids such as oleic acid are used in the preparation ofinjectables.

Pharmaceutically acceptable compositions described herein may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans. Topical application for the lower intestinal tract can beeffected in a rectal suppository formulation (see above) or in asuitable enema formulation. Topically-transdermal patches may also beused.

The amount of a compound described herein that may be combined with thecarrier materials to produce a composition in a single dosage form willvary depending upon the host treated and the particular mode ofadministration. In some embodiments, provided compositions should beformulated so that a dosage of between 0.01-100 mg/kg body weight/day ofthe inhibitor, such as e.g., 0.1-100 mg/kg body weight/day, can beadministered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of compound described herein in the composition will also dependupon the particular compound in the composition.

5. Uses of Compounds and Pharmaceutically Acceptable Compositions

In some embodiments, the compounds and compositions described herein areuseful in treating diseases and/or disorders associated withoverexpression of LSD1 and/or expression of a mutant form of LSD1, suchas those mutant forms that alter LSD1 substrate activity.

In some embodiments, the compounds and compositions described herein areuseful in treating diseases and/or disorders associated with cellularproliferation. In some embodiments, the compounds and compositionsdescribed herein are useful in treating diseases and/or disordersassociated with misregulation of cell cycle or DNA repair. In someembodiments, the compounds and compositions described herein are usefulin treating cancer. Exemplary types of cancer include breast cancer,prostate cancer, colon cancer, renal cell carcinoma, glioblastomamultiforme cancer, bladder cancer, melanoma, bronchial cancer, lymphomaand liver cancer.

In some embodiments, the present disclosure provides a method ofreducing the activity of LSD1 in a subject comprising the step ofadministering a compound described herein, or a composition comprisingany of the compounds herein. In some embodiments, the present disclosureprovides a method of reducing the activity of widetype-LSD1 in a subjectcomprising the step of administering a compound described herein, or acomposition comprising any of the foregoing. In some embodiments, thepresent disclosure provides a method of reducing the activity of amutant form of LSD1 in a subject comprising the step of administering acompound described herein, or a composition comprising any of theforegoing.

In some embodiments, the present disclosure provides a method oftreating a disease or condition related to cancer including e.g., tumorssuch as skin, breast, brain, cervical carcinomas, testicular carcinomas,etc. In one aspect, cancers that may be treated by the compositions andmethods described herein include, but are not limited to tumor typessuch as astrocytic, breast, cervical, colorectal, endometrial,esophageal, gastric, head and neck, hepatocellular, laryngeal, lung,oral, ovarian, prostate and thyroid carcinomas and sarcomas.

In some embodiments, the present disclosure provides a method oftreating a disease or condition selected from one or more of thefollowing, Cardiac: sarcoma (angiosarcoma, fibrosarcoma,rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma andteratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiatedsmall cell, undifferentiated large cell, adenocarcinoma), alveolar(bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma,chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus(squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma),stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductaladenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors,vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors,Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma),large bowel (adenocarcinoma, tubular adenoma, villous adenoma,hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma,Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra(squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma),prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma,embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma,interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors,lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone:osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibroushistiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma(reticulum cell sarcoma), multiple myeloma, malignant giant cell tumorchordoma, osteochronfroma (osteocartilaginous exostoses), benignchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma andgiant cell tumors; Nervous system: skull (osteoma, hemangioma,granuloma, xanthoma, osteitis deformans), meninges (meningioma,meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma,glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform,oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological:uterus (endometrial carcinoma), cervix (cervical carcinoma, pre-tumorcervical dysplasia), ovaries (ovarian carcinoma (serouscystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiedcarcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors,dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma,intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma),vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma(embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic:blood (myeloid leukemia (acute and chronic), acute lymphoblasticleukemia, chronic lymphocytic leukemia, myeloproliferative diseases,multiple myeloma, myelodysplastic syndrome), Hodgkin's disease,non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma,basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, molesdysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis;and Adrenal glands: neuroblastoma.

In one embodiment, the present disclosure provides a method of treatinga disease or condition seleted from CML, T-ALL, neuroblastoma, breastcancer, prostate cancer, herpes simplex virus reactivation, and HIVinfection comprising the step of administering to a subject in needthereof a compound described herein, or a pharmaceutically acceptablesalt thereof. In one alternative, the disease or condition is selectedfrom CML, T-ALL, and neuroblastoma.

EXEMPLIFICATION

The representative examples that follow are intended to help illustratethe present disclosure, and are not intended to, nor should they beconstrued to, limit the scope of what is described. Modifications andfurther embodiments, in addition to those shown and described herein,will become apparent to those skilled in the art.

It will further be appreciated that the present disclosure (includingthe Exemplification) contemplates individual the compounds describedherein. Where individual compounds exemplified are isolated and/orcharacterized as a salt, for example, as a trifluoroacetic acid salt,the present disclosure contemplates a free base of the salt, as well asother pharmaceutically acceptable salts of the free base.

Unless otherwise noted, all solvents, chemicals, and reagents wereobtained commercially and used without purification. The ¹H NMR spectrawere obtained in D₂O, CDCl₃, d₆-DMSO, CD₃OD, or d₆-acetone at 25° C. at400 MHz on an OXFORD (Varian) with chemical shift (δ, ppm) reportedrelative to TMS as an internal standard. HPLC-MS chromatograms andspectra were obtained with Shimadzu LC-MS-2020 system. Chiral analysisand purification were obtained with Yilite P270.

(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate

Step 1:

(E)-2-benzylidenebutanal

To a solution of benzaldehyde (130.0 kg, 1226.4 mol) in MeOH (1100.0 kg)was added NaOH (aq.) (10%, 500.0 kg). Then butanal (92.7 kg, 1287.5 mol)was added dropwise at 0° C. The reaction mixture was stirred at 10° C.for 10 h. The solvent was removed and the residue was acidified by HCl(aq., 4N) to pH=5. The mixture was extracted with EtOAc (500 kg+200 kg)and washed with brine (100 kg*2). The organic layer was concentratedunder vacuum. The crude product was distilled under vacuum (85˜95° C.,2-10 mmhg) to afford (E)-2-benzylidenebutanal (130.0 kg, GC>95%) as ayellowish oil.

Step 2:

ethyl (E)-4-((E)-benzylidene)hex-2-enoate

Ethyl 2-(diethoxyphosphoryl)acetate (182.0 kg, 812.5 mol) was dissolvedin THF (2200 kg) and cooled to 0° C. Potassium tert-butanolate (105.0kg, 937.5 mol) was added in one portion and the reaction mixture wasstirred vigorously at 0° C. for 15 min. (E)-2-benzylidenebutanal (130.0kg, 812.5 mol 1.0 eq) was added to the reaction mixture and the reactionmixture was stirred at 25° C. for 16 h. The reaction mixture wasquenched with H₂O (320.0 kg) and extracted with EtOAc (360.0 kg). Theorganic layer was dried over sodium sulfate and filtered. The solventwas evaporated under reduced pressure to afford crude product as an oil.The oil was dissolved in petroleum ether (560.0 kg), filtered through apad of silica gel (30.0 kg), and the filter cake was washed withpetroleum ether. The combined filtrate was concentrated afford ethyl(E)-4-((E)-benzylidene)hex-2-enoate as an oil (170.0 kg). This materialwas used in the next step without further purification.

Step 3:

ethyl (trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylate

Potassium tert-butanolate (103.5 kg, 924.1 mol) was dissolved in DMSO(1900 kg) under an inert atmosophere. The mixture was stirred for 15 minat 25° C. before addition of trimethylsulfoxonium iodide (210.0 kg,959.0 mol). This mixture was stirred for 45 min at 25° C. beforeaddition of ethyl (E)-4-((E)-benzylidene)hex-2-enoate (170.0 kg, 738.0mol). The reaction was warmed to 50˜55° C. and stirred for 16 h. Thereaction was quenched with water (600 kg). The mixture was extractedwith petroleum ether (800 kg). The petroleum ether phase was evaporatedunder reduced pressure to afford ethyl(trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylate (76.5kg) as brown liquid. This material was used in the next step withoutfurther purification

Step 4:

(trans)-2-(((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylic acid

Ethyl (trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylate(76.5 kg kg, 313.1 mol) was dissolved in water (320 kg) and methanol(360 kg). To this solution was added sodium hydroxide (41.0 kg, 1025mol) and the solution was stirred for 3 h at 50° C. The mixture wasconcentrated to about 300 L and acidified to pH=4 using HCl (aq., 4N).This solution was extracted with EtOAc (520 kg) and the layersseparated. The organic layer was washed with brine, dried over sodiumsulfate, filtered, and concentrated to afford(trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylic acid(62.5 kg). This material was used in next step without furtherpurification.

Step 5:

(trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine hydrochloride

To a solution of(trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropane-1-carboxylic acid(26.0 kg, 120.4 mol) in toluene (270 kg) were added triethylamine (36.3kg, 359 mol) and diphenylphosphoryl azide (38.0 kg, 138.1 mol) at 0° C.The reaction mixture was stirred at 0˜10° C. for 3 h. Water (100 kg) wasadded and the mixture was extracted with EtOAc (80.0 kg) and the organiclayer was concentrated under vacuum to afford a crude oil. The crude oilwas dissolved in petroleum ether (50 kg). The suspension was filteredthrough a pad of silica gel and the filter cake was washed withpetroleum ether. The combined filtrates were concentrated to affordcrude acyl azide. The crude acyl azide was taken up in toluene (100 kg)and the solution was heated to 85° C. for 2 h. The organic layer wasconcentrated to ˜60 L. Potassium trimethylsilanolate (23.0 kg, 177.9mol) was added to the toluene mixture at room temperature and stirredfor 1.5 h. The reaction was then treated with HCl (2N aq., 120 kg) andstirred. The layers were separated. The organic layer was extractedtwice with HCl (2N aq., 20 kg) and the aqueous extracts were combined.The aqueous layer was extracted with MTBE (30 kg) once, and thenbasified to pH=10-11 with sodium hydroxide (47 kg, 30% aqueous). Theaqueous layer was then extracted with MTBE (150 kg). The organicextracts were washed with brine, dried with sodium sulfate and filtered.To this MTBE solution was added HCl (23.5 kg, 12% in diethyl ether) anda white solid precipitated from solution. The solid was collected toafford (trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-aminehydrochloride (16.5 kg, 73.8 mol).

Step 6:

(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate

(trans)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine hydrochloride(16.5 kg, 73.8 mol) was charged to a reacter and K₃PO₃ (46 L, 37 wt %)was added followed by EtOAc (150 kg). The reaction mixture was stirredat 0˜10° C. for 3 h. The organic layer was separated and concentrated todryness to afford an oil. S-Mandelic acid (8.4 kg, 55.35 mol) and 95%EtOH (165 kg) were added to the reactor and the reaction was stirred atroom temperature for 72 h (until ee>99%). The solid was collected viafiltration to afford(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate (5.8 kg, 17.1 mol). LCMS m/z 188 [M+H]⁺.The absolute stereochemistry was determined by X-ray crystallography.See FIG. 1.

N-(2,2-dimethoxyethyl)methanesulfonamide

To a solution of 2,2-dimethoxyethanamine (518 μL, 4.75 mmol) indichloromethane (4.75 mL) at 0° C. was added triethylamine (1.32 mL,9.50 mmol) followed by methanesulfonyl chloride (550 μL, 7.12 mmol). Themixture was stirred at 0° C. for 30 minutes. The reaction was dilutedwith dichloromethane and washed with saturated aqueous sodiumbicarbonate. The aqueous layer was extracted twice with dichloromethane.The combined organic layers were dried with sodium sulfate thenconcentrated under reduced pressure to afford 1.18 g of crude compoundas light yellow oil. The crude mixture was then diluted in acetone:water(1:1, 4 mL) and treated with amberlyst-15 hydrogen form (1.2 g). Theresulting mixture was stirred for 2 hours at room temperature. Thesuspension was filtered through celite and concentrated under reducedpressure to afford crude N-(2,2-dimethoxyethyl)methanesulfonamide (900mg) as a yellow oil. The product was used without further purification.¹H NMR (400 MHz, dmso) δ 7.13 (t, J=6.0 Hz, 1H), 4.36 (t, J=5.5 Hz, 1H),3.28 (s, 6H), 3.02 (t, J=5.7 Hz, 2H), 2.90 (s, 3H).

N-(2-oxoethyl)ethanesulfonamide

Step 1:

N-(2,2-dimethoxyethyl)ethanesulfonamide

To a solution of 2,2-dimethoxyethanamine (247 μL, 2.28 mmol) in DCM (1.9mL) at 0° C. was added triethylamine (634 μL, 4.56 mmol) followed byethanesulfonyl chloride (212 μL, 2.28 mmol). The mixture was stirred at0° C. for 30 min. The reaction was diluted with DCM and washed with sat.NaHCO₃. The aqueous layer was back-extracted twice with DCM. Thecombined organic layers were dried with Na₂SO₄ and then concentratedunder reduced pressure to afford crudeN-(2,2-dimethoxyethyl)ethanesulfonamide as a brown oil.

Step 2:

N-(2-oxoethyl)ethanesulfonamide

To a solution of crude N-(2,2-dimethoxyethyl)ethanesulfonamide inacetone: water (1:1, 4 mL) was added Amberlyst-15 Hydrogen form (1.0 g).The resulting mixture was stirred over two days at room temperature. Thesuspension was filtered through celite and concentrated under reducedpressure to afford crude N-(2-oxoethyl)ethanesulfonamide (614 mg) as ayellow oil. Used without further purification. 1H NMR (400 MHz, CDCl₃) δ9.68 (s, J=1.1 Hz, 1H), 4.15 (d, J=5.2 Hz, 2H), 3.07 (q, J=7.4 Hz, 2H),1.40 (t, J=7.4 Hz, 3H).

Using the appropriate starting material and modifications the followingintermediates in Table 1 were synthesized using the synthetic proceduresdescribed for N-(2-oxoethyl)ethanesulfonamide.

TABLE 1 Structure Name

N-(2-oxoethyl)acetamide

N-(2-oxoethyl)cyclopropanesulfonamide

N-(2-oxoethyl)propane-2-sulfonamide

2-methoxy-N-(2-oxoethyl)ethane-1-sulfonamide

2,2-dimethyl-1-(methylsulfonyl)aziridine

To a solution of 2-amino-2-methylpropan-1-ol (1.0 g, 11.2 mmol) in DCM(22.4 mL) at 0° C. was added triethylamine (7.79 mL, 56.0 mmol) followedby methanesulfonyl chloride (2.16 mL, 28.0 mmol). The mixture wasstirred at 0° C. for 30 min and was then allowed to warm to roomtemperature overnight. The reaction was diluted with DCM and washed withsat. NaHCO₃. The aqueous layer was extracted once with DCM. The combinedorganic layers were dried over Na₂SO₄ and then concentrated underreduced pressure to afford crude2,2-dimethyl-1-(methylsulfonyl)aziridine (1.95 g) compound as a brownoil.

2,2,2-trifluoro-N-((1R,2S)-2-(E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(7-azaspiro[3.5]nonan-2-yl)acetamidehydrochloride

Step 1:

tert-butyl2-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonane-7-carboxylate

To a suspension of (1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropanamine(S)-2-hydroxy-2-phenylacetate (3.5 g, 10.3 mmol) in 1,2-dichloroethane(75 mL) at 0° C. was added tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (2.58 g, 10.8 mmol) followedby sodium triacetoxyborohydride (4.78 g, 22.6 mmol). The reactionmixture was allowed to warm to 23° C. and stirred for 4 hours.Additional tert-butyl 2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (123 mg,0.51 mmol) and sodium triacetoxyborohydride (328 mg, 1.55 mmol) werethen added and the reaction mixture was stirred at 23° C. for 2 hours.The reaction mixture was then diluted with a saturated aqueous K₂CO₃solution (50 mL) and extracted with DCM (3×20 mL). The combined organiclayers were washed with brine (20 mL), dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The crude residue was purified viaBiotage (gradient 0 to 100% EtOAc in hexanes) to afford tert-butyl2-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonane-7-carboxylate(3.235 g). LCMS (ESI) m/z 411.3 [M+H]⁺.

Step 2:

tert-butyl2-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-7-azaspiro[3.5]nonane-7-carboxylate

To a solution of tert-butyl2-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonane-7-carboxylate(3.226 g, 7.84 mmol) in DCM (50 mL) at 0° C. was addedN,N-diisopropylethylamine (2.03 mL, 11.7 mmol) followed bytrifluoroacetic anhydride (1.40 mL, 10.1 mmol). The reaction mixture wasallowed to warm to 23° C. and stirred for 4 hours. The reaction mixturewas then diluted with a saturated aqueous NaHCO₃ solution (30 mL) andthe two layers were separated. The organic layer was washed with asaturated aqueous NH₄Cl solution (2×20 mL), dried over Na₂SO₄, filtered,and concentrated under reduced pressure. The crude residue was purifiedvia Biotage (gradient 0 to 50% EtOAc in hexanes) to afford tert-butyl2-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-7-azaspiro[3.5]nonane-7-carboxylate(3.258 g). LCMS (ESI) m/z: 529.2 [M+Na]⁺.

Step 3:

2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(7-azaspiro[3.5]nonan-2-yl)acetamidehydrochloride

To a solution of tert-butyl2-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-7-azaspiro[3.5]nonane-7-carboxylate(3.254 g, 6.41 mmol) in dioxane (32 mL) at 23° C. was added a 4.0 Msolution of HCl in dioxane (12.8 mL, 51.2 mmol) dropwise. The reactionmixture was stirred at 23° C. for 2 hours. Additional HCl (12.8 mL, 51.2mmol, 4.0 M solution in dioxane) was then added and the reaction mixturewas stirred at 23° C. for 2 hours. The reaction mixture was thenconcentrated under reduced pressure to afford2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(7-azaspiro[3.5]nonan-2-yl)acetamidehydrochloride as a crude yellow foam (3.185 g). LCMS (ESI) m/z: 407.2[M+H]⁺.

2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate

Step 1:

tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate

To (1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropanamine(S)-2-hydroxy-2-phenylacetate (8.65 g, 25.5 mmol) and tert-butyl6-oxo-2-azaspiro[3.3]heptane-2-carboxylate (5.15 g, 24.3 mmol) in1,2-DCE (100 mL) was added sodium triacetoxyborohydride (10.8 g, 51mmol). After 30 min. the reaction was quenched with K₂CO₃ (aq.) andextracted with DCM (2×150 mL). The organic phase was concentrated andthe crude residue was purified via column chromatography (50 g column,5% to 100% EtOAc:hexanes) to afford tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate(4.61 g). LCMS m/z: 383.7 [M+H]⁺.

Step 2:

tert-butyl6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate

To tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate(4.61 g, 12 mmol) dissolved in DCM was added diisopropylethylamine (2.81mL, 16.2 mmol). The solution was cooled to 0° C. before addition oftrifluoroacetic anhydride (2.08 mL, 15 mmol). The reaction mixture wasstirred for 2 h, while warming to room temperature. The volatiles wereevaporated under reduced pressure to afford crude tert-butyl6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate.LCMS m/z 479 [M+H]⁺.

Step 3:

2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate

Tert-butyl 6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate(5.74 g, 12 mmol) was dissolved in DCM (80 mL) and cooled to 10° C.before addition of trifluoroacetic acid (20 mL, 240 mmol). The solutionwas allowed to warm to room temperature and stirred for 3 h. Thereaction was concentrated to afford crude2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate. LCMS m/z 379.2 [M+H]⁺.

Compound 1:2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanol

Step 1:

2,2,2-trifluoro-N-(2-(2-hydroxyethyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide

A solution of2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate (7.57 g, 15.3 mmol) and glycoaldehyde dimer (5.51g, 45.9 mmol) in methanol was treated with acetic acid (2.0 mL) andsodium cyanoborohydride (2.88 g, 45.9 mmol). The reaction was heated to60° C. and monitored by LCMS. After 1 hour, the reaction was quenchedwith 10% aqueous potassium carbonate to a pH of 8. Volatiles were thenremoved under vacuum. The crude mixture was extracted twice with2-methyltetrahydrofuran. The organic layer was then dried with sodiumsulfate, filtered and volatiles were removed under vacuum. The desiredproduct2,2,2-trifluoro-N-(2-(2-hydroxyethyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamidewas used without further purification. LCMS (ESI) m/z: 423.1 [M+H]⁺.

Step 2:

2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanolbis(2,2,2-trifluoroacetate)

To a round bottom flask,2,2,2-trifluoro-N-(2-(2-hydroxyethyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(6.46 g, 15.3 mmol) was dissolved in methanol (75 mL), cooled to 0° C.and treated with 10 mL of 10% aqueous sodium hydroxide. The reaction wasstirred under nitrogen atmosphere for 30 minutes and was quenched with2,2,2-trifluoroacetic acid to a pH of 2.0. Volatiles were removed undervacuum. The desired product was purified by reverse phase columnchromatography using 10-30% CH₃CN/0.1% aqueous 2,2,2-trifluoroaceticacid. Pure fractions were combined, frozen and lyophilized to afford2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanolbis-(2,2,2-trifluoroacetic acid) (2.90 g). LCMS (ESI) m/z: 327.3 [M+H]⁺.¹H NMR (400 MHz, DMSO-d₆) δ 9.87 (br s, 1H), 9.21 (br s, 2H), 7.39-7.31(m, 2H), 7.27-7.17 (m, 3H), 6.21 (s, 1H), 5.23 (br s, 1H), 4.25-4.11 (m,2H), 4.11-4.06 (m, 1H), 3.98 (m, 1H), 3.85-3.69 (m, 1H), 3.55 (s, 2H),3.25-3.11 (m, 2H), 2.80-2.72 (m, 1H), 2.72-2.65 (m, 1H), 2.65-2.53 (m,1H), 2.48 -2.35 (m, 2H), 2.30-21.8 (m, 2H), 2.02-1.90 (m, 1H), 1.22-1.14(m, 2H), 1.12 (t, J=7.6 Hz, 3H).

Compound 2:2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol

Step 1:

N-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide

To a round bottom flask,2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate (2.75 g, 5.58 mmol) dissolved in methanol (18.6mL) was added 1,3-dihydroxypropan-2-one (3.0 g, 33.4 mmol) and aceticacid (10 drops). The solution was then treated with sodiumcyanoborohydride (2.09 g, 33.4 mmol) and stirred for 1 h. The reactionwas then quenched to pH 8 using 10% potassium carbonate aqueous andvolatiles were removed under vacuum. The crude mixture was extractedtwice with 2-methyltetrahydrofuran. The combined organic layers werewashed once with brine, then dried with sodium sulfate, filtered andconcentrated to afford crudeN-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide.The crude product was used without further purification. LCMS (ESI) m/z:453.4 [M+H]⁺.

Step 2:

2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diolbis(2,2,2-trifluoroacetate)

N-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(2.52 g, 5.58 mmol) was dissolved in methanol, cooled in an ice bath andtreated with 10% aqueous NaOH and allowed to reach room temperature.After 30 min, the reaction was quenched to pH 2 using TFA, and volatileswere removed under reduced pressure. The crude mixture was purified byreverse phase column chromatography (60 g) using 10-30% MeCN/0.1% TFAaq. as eluent. Fractions were combined, frozen and lyophilized to afford2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diolbis(2,2,2-trifluoroacetate) as a white amorphous solid (740 mg). LCMS(ESI) m/z: 357.4 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 10.00 (s br, 1H),9.33 (s br, 2H), 7.42-7.31 (m, 2H), 7.28-7.16 (m, 3H), 6.20 (s, 1H),5.28 (s br, 2H), 4.30 (t, J=15.6 Hz, 2H), 4.14 (s, 1H), 3.99 (s, 1H),3.86-3.68 (m, 1H), 3.58 (dd, J=12.7, 9.6 Hz, 4H), 3.29 (s, 2H), 2.72 (s,2H), 2.40 (d, J=8.6 Hz, 1H), 2.31-2.15 (m, 2H), 1.97 (s, 1H), 1.15 (dt,J=15.1, 5.9 Hz, 5H).

Scale Up and Formation of Monocitrate Salt of2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol

Step 1:

tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate

Charge (1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate (160 g, 1.0 eq.) into a 3 L flask anddissolved in DCM (1000 mL). To this flask was added a solution of 10%K₂CO₃ (500 mL) and water (500 mL). The solution was stirred for 10 min.The layers were separated and the aqueous layer was extracted with DCM(2×, 300 mL). The combined organics layer was dried over Na₂SO₄ (100 g),filtered, and concentrated to afford(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine as a lightyellow oil. This material was dissolved in 2-MeTHF (500 mL) andconcentrated. This material was then dissolved in 2-MeTHF (1000 mL) andtransferred to a 10 L flask. To the 10 L flask was added 2-MeTHF (1000mL) to completely dissolved the freebase and to this solution was addedtert-butyl 6-oxo-2-azaspiro[3.3]heptane-2-carboxylate (100 g). Thereaction was stirred for 10 min before addition of NaBH(OAc)₃ (200 g,2.0 eq). The reaction was stirred at room temperature for 4 h.Additional NaBH(OAc)₃ (8 g, 0.08 eq) was added and the solution wasstirred at room temperature for 2 h. This solution was basified to pH=8via addition of a 10% K₂CO₃ solution (950 mL) and the reaction wasstirred for 30 min at room temperature. The layers were separated andthe water layer was extracted with DCM (1×, 500 mL). The combinedorganics layer was concentrated (bath temperature<45° C.) to affordcrude tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate(220 g).

Step 2:

tert-butyl6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate

To a flask charged with crude tert-butyl6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptane-2-carboxylate(200 g) was added DCM (2000 mL) and Hunig's base (100 mL). The reactionwas cooled to 0° C. before addition of trifluoroacetic anhydride (120 g)dropwise over 30 min. The reaction was stirred at 0° C. before warmingto 10-20° C. and stirring for 16 h. Water (2000 mL) was added to thisreaction and the layers were separated. The organics layer wasconcentrated to afford crude product (264 g). The crude oil wasdissolved in heptane (500 mL) and stirred for 30 min to afford asuspension. This material was then purified via silica gelchromatography (1.5 kg, 100-200 mech silica gel; gradient: 1:20 ethylacetate:heptane; 1:10 ethyl acetate heptane; 1:5 ethyl acetate:heptane)to obtain tert-butyl6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate(Fraction A: 114 g, 96% pure; Fraction B: 72 g, 86% pure).

Step 3:

2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate

To a flask charged with tert-butyl6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptane-2-carboxylate(104 g) was added DCM (1050 mL) and the solution was cooled to 0° C. Tothis solution was added TFA (240 g, 10 eq.) and the solution was warmedto room temperature. The reaction was stirred for 2 h before additionalTFA (40 g, 2.0 eq.) was added. The reaction was stirred for 1 h beforebeing concentrated to afford a brown oil. The brown oil was dissolved inMeOH (500 mL) and concentrated. This step was repeated (3×) to affordcrude2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate (365.7 g).

Step 4:

N-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide

To a flask charged with crude2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate (365.7 g, 1.0 eq.) was added MeOH (1500 mL). Tothis solution was added 1,3-dihydroxypropan-2-one (120 g, 1.33 eq.) andthe solution was stirred a room temperature for 20 min before additionof AcOH (20 mL, 1.41 eq.). The reaction was stirred for 10 min beforeaddition of NaBH₃CN (84 g, 6.01 eq.). The reaction was placed in a coldbath to keep the temperature below 40° C. The reaction was stirred for 1h and during the course of the reaction the mixture turned light yellow.This solution was basified to pH=8 via addition of 10% K₂CO₃ solution(400 mL). The mixture was concentrated to remove the MeOH (bathtemperature<45° C.). The resulting solution was dissolved in DCM (500mL) and water (200 mL) was added. The layers were separated and theaqueous layer was extracted with DCM (2×). The combined organics layerswere dried over Na₂SO₄, filtered, and concentrated to afford crudeN-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(110.6 g).

Step 5:

2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol

To a flask charged with crudeN-(2-(1,3-dihydroxypropan-2-yl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(110.6 g) was added 2-MeTHF (1100 mL). To this mixture was slowly addedNaOH solution (20 g NaOH dissolved in 500 mL water). The reaction wasstirred at room temperature for 1 h. The layers were separated and theaqueous layer was extracted with 2-MeTHF (2×). The combined organicslayers were washed with brine, dried over Na₂SO₄, filtered, andconcentrated to afford a yellow oil (190 g). The yellow oil wasdissolved in ethyl acetate and washed with brine. The organics layer wasconcentrated (bath temperature<45° C.) to afford2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol(76.4 g) as a light yellow solid.

Step 6:

2-(6-(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol2-hydroxypropane-1,2,3-tricarboxylate

To a flask charged with2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol(50 g, 1.0 eq.) was added a mixture of DCM:MeOH (500 mL:40 mL). Thissolution was stirred at room temperature for 10 min before dropwiseaddition of citric acid (40 g, 1.43 eq) dissolved in MeOH (90 mL). Themixture was stirred at room temperature for 1 h before the precipitatewas collected via filtration. The solid was washed with a mixture ofMeOH:DCM (3:20, 200 mL). The filter cake was dried under vacuum at 40°C. for 16 h to afford2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3-diol2-hydroxypropane-1,2,3-tricarboxylate (77.4 g) as a crystalline product.1H NMR (400 MHz, D₂O) δ=7.46-7.33 (m, 2H), 7.31-7.25 (m, 3H), 6.23 (s,1H), 4.40 (s, 2H), 4.32 (br s, 2H), 3.93 (quin, J=8.1 Hz, 1H), 3.84-3.71(m, 4H), 3.40 (quin, J=4.6 Hz, 1H), 2.90-2.52 (m, 9H), 2.36 -2.19 (m,2H), 2.09-2.00 (m, 1H), 1.30-1.19 (m, 2H), 1.11 (t, J=7.8 Hz, 3H). Thestructure was confirmed by X-ray crystallography. See FIG. 2.

Using the appropriate starting material and modifications, the followingcompounds in Table 2 were synthesized using the synthetic proceduresdescribed for Compound 1. Only the 1R,2S isomers are shown in the tablein an effort to reduce page length.

TABLE 2 LCMS Compound Structure/name 1H NMR m/z 3

¹H NMR (400 MHz, DMSO-d₆) δ 10.43 (s, 3H), 9.25 (s, 13H), 7.98 (s, 17H),7.33 (t, J = 7.5 Hz, 15H), 7.28-7.11 (m, 22H), 6.18 (s, 7H), 4.08 (s,20H), 3.77 (s, 9H), 2.93 (s, 19H), 2.71 (s, 13H), 2.58 (s, 14H),2.44-2.35 (m, 12H), 2.30-2.16 (m, 17H), 1.95 (s, 8H), 1.25-1.06 (m,38H). 326.4 [M + H]⁺ 4

1H NMR (400 MHz, DMSO-d6) δ 9.91 (br s, 1H), 9.22 (br s, 2H), 7.35 (t, J= 7.5 Hz, 2H), 7.30 (t, J = 6.0 Hz, 1H), 7.26-7.17 (m, 3H), 6.21 (s,1H), 4.28-4.19 (m, 1H), 4.19-4.06 (m, 3H), 3.88-3.70 (m, 1H), 3.28-3.20(m, 2H), 3.20- 3.11 (m, 2H), 2.96 (s, 3H). 2.82- 2.64 (m, 2H), 2.63-2.52(m, 1H), 2.47-2.35 (m, 1H), 2.29- 2.17 (m, 2H), 2.02-1.89 (m, 1H), 1.17(t, J =7.3 Hz, 2H), 1.12 (t, J = 7.5 Hz, 3H). 404.1 [M + H]⁺ 5

1H NMR (400 MHz, DMSO-d6) δ 9.81 (br s, 1H), 9.21 (br s, 2H), 8.05 (t, J= 5.6 Hz, 1H), 7.39- 7.31 (m, 2H), 7.27-7.17 (m, 3H), 6.21 (s, 1H),4.27-4.18 (m, 1H), 4.15-4.05 (m, 3H), 3.85-3.74 (m, 1H), 3.26-3.10 (m,4H), 2.80- 2.70 (m, 1H), 2.70-2.63 (m, 1H), 2.63-2.54 (m, 1H), 2.47-2.36 (m, 1H), 2.30-2.18 (m, 2H), 2.00-1.91 (m, 1H), 1.83 (s, 3H),1.21-1.14 (m, 2H), 1.12 (t, J = 7.5 Hz, 3H). 368.4 [M + H]⁺ 6

1H NMR (400 MHz, DMSO-d6) δ 9.91 (br s, 1H), 9.20 (br s, 2H), 7.42-7.31(m, 3H), 7.28-7.16 (m, 3H), 6.21 (s, 1H), 4.27-4.19 (m, 1H), 4.19-4.15(m, 1H), 4.15- 4.06 (m, 2H), 3.87-3.72 (m, 1H), 3.27-3.14 (m, 4H), 2.81-2.66 (m, 2H), 2.66-2.60 (m, 1H), 2.60-2.52 (m, 1H), 2.47-2.35 (m, 1H),2.30-2.17 (m, 2H), 2.01- 1.91 (m, 1H), 1.21-1.15 (m, 2H), 1.12 (t, J =7.5 Hz, 3H), 1.04- 0.95 (m, 2H), 0.95-0.87 (m, 2H). 430.4 [M + H]⁺ 7

1H NMR (400 MHz, DMSO-d6) δ 9.92 (br s, 1H), 9.22 (br s, 2H), 7.40-7.29(m, 3H), 7.29-7.15 (m, 3H), 6.21 (s, 1H), 4.28-4.19 (m, 1H), 4.19-4.03(m, 3H), 3.79 (br s, 1H), 3.26-3.17 (m, 2H), 3.16-3.10 (m, 2H), 3.07 (q,J = 7.4 Hz, 2H), 2.80-2.64 (m, 2H), 2.64-2.52 (m, 1H), 2.47-2.35 (m,1H), 2.31-2.17 (m, 2H), 2.03- 1.90 (m, 1H), 1.19 (t, J = 7.4 Hz, 3H),1.17-1.15 (m, 2H), 1.12 (t, J = 7.5 Hz, 3H). 418.1 [M + H]⁺ 8

1H NMR (400 MHz, DMSO-d6) δ 9.93 (br s, 1H), 9.25 (br s, 2H), 7.40-7.28(m, 3H), 7.27-7.15 (m, 3H), 6.21 (s, 1H), 4.28-4.20 (m, 1H), 4.20-4.06(m, 3H), 3.86- 3.73 (m, 1H), 3.26 (dd, J = 13.4, 6.6 Hz, 1H), 3.23-3.09(m, 4H), 2.80-2.64 (m, 2H), 2.64-2.53 (m, 1H), 2.48-2.36 (m, 1H), 2.32-2.17 (m, 2H), 2.03-1.92 (m, 1H), 1.22 (d, J = 6.8 Hz, 6H), 1.20 1.15 (m,J = 12.4, 5.3 Hz, 2H), 1.12 (t, J = 7.5 Hz, 3H). 432.2 [M + H]⁺ 9

1H NMR (400 MHz, DMSO-d6) δ 9.83 (br s, 1H), 9.15 (brs, 2H), 7.39-7.33(m, 2H), 7.31 (t, J = 6.1 Hz, 1H), 7.27-7.17 (m, 3H), 6.21 (s, 1H),4.29-4.19 (m, 1H), 4.19-4.06 (m, 3H), 3.87-3.72 (m, 1H), 3.65 (t, J =6.0 Hz, 2H), 3.51-3.42 (m, 1H), 3.27 (s, 3H), 3.21 (s, 2H), 3.18-3.08(m, 2H), 2.81-2.64 (m, 2H), 2.64-2.53 (m, 1H), 2.46-2.35 (m, 1H), 2.29-2.18 (m, 2H), 2.01-1.90 (m, 1H), 1.21-1.15 (m, 2H), 1.12 (t, J = 7.5 Hz,3H). 448.3 [M + H]⁺ 10

¹H NMR (400 MHz, DMSO-d₆) δ 10.01-9.78 (m, 2H), 9.59 (br s, 1H),7.37-7.31 (m, 2H), 7.25- 7.18 (m, 3H), 6.20 (s, 1H), 5.39 (br s, 2H),3.89-3.81 (m, 1H), 3.77 (d, J = 4.5 Hz, 4H), 3.44-3.39 (m, 2H),3.23-3.15 (m, 2H), 3.13- 3.05 (m, 1H), 2.69-2.61 (m, 1H), 2.38-2.31 (m,1H), 2.29-2.20 (m, 3H), 2.19-2.09 (m, 3H), 1.99- 1.86 (m, 4H), 1.36-1.27(m, 1H), 1.17-1.09 (m, 4H). 385.5 [M + H]⁺ 11

(500 MHz, DMSO-d6) δ 9.89 (br s, 1H), 9.84 (br s, 1H), 9.79 (br s, 1H),7.37-7.32 (m, 2H), 7.25- 7.19 (m, 3H), 6.20 (s, 1H), 5.31 (br s, 1H),3.89-3.82 (m, 1H), 3.78- 3.72 (m, 2H), 3.43-3.37 (m, 2H), 3.14-3.06 (m,2H), 3.02-2.82 (m, 2H), 2.69-2.62 (m, 1H), 2.38- 2.30 (m, 1H), 2.29-2.20(m, 3H), 2.17-2.09 (m, 3H), 1.97- 1.81 (m, 4H), 1.34-1.28 (m, 1H),1.17-1.09 (m, 4H) 355.3 [M + H]⁺

Compound 12:2-methyl-1-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propan-2-olbis(2,2,2-trifluoroacetate)

Step 1:

2,2,2-trifluoro-N-(2-(2-hydroxy-2-methylpropyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide

To a sealed tube was added2,2,2-trifluoro-N-((1R,2S)-2-(E)-1-phenylbut-1-en-2-yl)cyclopropyl)-N-(2-azaspiro[3.3]heptan-6-yl)acetamide2,2,2-trifluoroacetate (200 mg, 406 μmol), 2,2-dimethyloxirane (54.0 μL,609 μmol), diisopropylethylamine (210 μL, 1.21 mmol) and EtOH (4.1 mL).The reaction mixture was stirred at 50° C. for 18 h. The mixture wasconcentrated under reduced pressure, diluted with ethyl acetate thenquenched with 1N HCl. The layers were separated and the aqueous wasextracted once with ethyl acetate. The combined organic layer was washedonce with brine, dried with sodium sulfate and evaporated under reducedpressure to afford2,2,2-trifluoro-N-(2-(2-hydroxy-2-methylpropyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamideyellow oil (296 mg). LCMS m/z 451.3 [M+H]⁺.

Step 2:

2-methyl-1-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propan-2-olbis(2,2,2-trifluoroacetate)

To a solution of2,2,2-trifluoro-N-(2-(2-hydroxy-2-methylpropyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(250 mg, 554 μmol) in MeOH (2.8 mL) at room temperature was added NaOH(1M, 2.77 mL, 2.77 mmol). The mixture was allowed to stir at roomtemperature for 60 min. The mixture was quenched with TFA thenconcentrated under reduced pressure. The crude product was purified on a30 g C-18 column using McCN/0.1% TFA H2O as eluent with the followinggradient (2 CV 0%, 20 CV 0-30%, 3 CV 30%) to afford2-methyl-1-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propan-2-olbis(2,2,2-trifluoroacetate) (22.0 mg) after lyophilisation. LCMS m/z355.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 9.59 (br s, 1H), 9.27 (br s,2H), 7.35 (t, J=7.6 Hz, 2H), 7.29-7.14 (m, 3H), 6.20 (s, 1H), 5.10 (brs, 1H), 4.37-4.25 (m, 1H), 4.25-4.07 (m, 3H), 3.83-3.64 (m, 1H),3.18-3.09 (m, 2H), 2.79-2.65 (m, 2H), 2.65-2.52 (m, 1H), 2.47-2.33 (m,1H), 2.30-2.13 (m, 2H), 2.03-1.89 (m, 1H), 1.23-1.15 (m, 2H), 1.13 (s,6H), 1.11 (t, J=7.6 Hz, 3H).

Using the appropriate starting material and modifications, the followingexamples in Table 3 were synthesized using the synthetic proceduresdescribed for Compound 12. Only the 1R,2S isomers are shown in the tablein an effort to reduce page length.

TABLE 3 LCMS Compound Structure/name 1H NMR m/z 13

1H NMR (400 MHz, DMSO-d6) δ 10.5 (s br, 1H), 9.3 (s br, 2H), 8.46 (s br,1H), 7.4-7.3 (m, 2H), 7.3- 7.2 (m, 3H), 6.20 (s, 1H), 4.79 (s, 1H), 4.23(s, 1H), 4.2-4.0 (m, 3H), 3.96 (m, 2H), 3.76 (s, 1H), 3.15 (2H),2.72-2.68 (m, 2H), 2.58 (m, 1H), 2.43 (m, 1H), 2.24 (m, 2H), 1.96 (m,1H), 1.12 (m, 5H) 384.2 [M + H]⁺ 14

1H NMR (400 MHz, DMSO-d6) δ 10.51 (s br, 1H), 9.44 (s br, 2H), 8.79 (sbr, 1H), 7.75 (d, J = 59.3 Hz, 2H), 7.35 (d, J = 7.6 Hz, 2H), 7.28-7.14(m, 3H), 6.20 (s, 1H), 4.20 (d, J = 14.2 Hz, 1H), 4.04 (dd, J = 12.5,6.9 Hz, 1H), 3.92 (dd, J = 12.1, 6.4 Hz, 1H), 3.76 (s, 1H), 2.72 (s,1H), 2.58 (ddd, J = 16.3, 12.1, 6.0 Hz, 2H), 2.47-2.35 (m, 1H),2.30-2.16 (m, 2H), 1.98 (s, 1H), 1.42 (s, 6H), 1.29-1.05 (m, 5H). 368.7[M + H]⁺ 15

1H NMR (400 MHz, DMSO-d6) δ 9.54 (s, 1H), 9.18 (br s, 2H), 7.39- 7.32(m, 2H), 7.27-7.17 (m, 3H), 7.15 (s, 1H), 6.21 (s, 1H), 4.41- 4.30 (m,1H), 4.30-4.17 (m, 3H), 3.87-3.70 (m, 1H), 3.20 (s, 2H), 3.03 (s, 3H),2.81-2.65 (m, 3H), 2.64-2.52 (m, 1H), 2.48-2.35 (m, 2H), 2.31-2.16 (m,2H), 1.97 (s, 1H), 1.29 (s, 3H), 1.21-1.15 (m, 2H), 1.12 (t, J = 7.5 Hz,3H). 432.2 [M + H]⁺ 16

(400 MHz, DMSO-d6) δ 9.99- 9.76 (m, 2H), 9.52 (br s, 1H), 7.37- 7.32 (m,2H), 7.25-7.18 (m, 3H), 6.20 (s, 1H), 5.46 (d, J = 4.2 Hz, 1H),4.17-4.08 (m, 1H), 3.89- 3.80 (m, 1H), 3.44-3.37 (m, 2H), 3.09-3.01 (m,1H), 2.99-2.84 (m, 3H), 2.69-2.62 (m, 1H), 2.37- 2.29 (m, 1H), 2.29-2.20(m, 3H), 2.18-2.09 (m, 3H), 1.96-1.83 (m, 4H), 1.35-1.27 (m, 1H), 1.16-1.07 (m, 7H) 369.3 [M + H]⁺ 17

(400 MHz, DMSO-d6) δ 10.02 (br s, 1H), 9.95-9.79 (m, 2H), 7.37- 7.31 (m,2H), 7.25-7.18 (m, 3H), 6.20 (s, 1H), 3.89-3.81 (m, 1H), 3.69 (t, J =4.9 Hz, 2H), 3.37-3.31 (m, 2H), 3.30 (s, 3H), 3.25-3.18 (m, 2H),3.03-2.92 (m, 1H), 2.92- 2.80 (m, 1H), 2.70-2.61 (m, 1H), 2.37-2.29 (m,1H), 2.29-2.19 (m, 3H), 2.19-2.08 (m, 3H), 1.97- 1.80 (m, 4H), 1.35-1.27(m, 1H), 1.17-1.09 (m, 4H) 369.3 [M + H]⁺ 18

(400 MHz, DMSO-d6) δ 9.96- 9.82 (m, 2H), 9.11 (br s, 1H), 7.38- 7.32 (m,2H), 7.25-7.18 (m, 3H), 6.20 (s, 1H), 5.26 (s, 1H), 3.89- 3.81 (m, 1H),3.45-3.38 (m, 2H), 3.09-3.03 (m, 3H), 3.01-2.94 (m, 1H), 2.69-2.62 (m,1H), 2.35- 2.21 (m, 4H), 2.18-2.10 (m, 3H), 2.05-1.94 (m, 2H), 1.92-1.82(m, 2H), 1.35-1.27 (m, 1H), 1.25 (s, 6H), 1.17-1.10 (m, 4H) 383.3 [M +H]⁺ 19

1H NMR (400 MHz, DMSO-d6) δ 9.90 (br. s., 2 H), 9.58 (br. s., 1 H),7.43-7.14 (m, 5 H), 6.19 (s, 1 H), 5.54 (br. s., 1 H), 4.98 (br. s., 1H), 4.03-3.77 (m, 2 H), 3.16 (d, J = 7.0 Hz, 1 H), 2.95 (dd, J = 9.8,13.5 Hz, 3 H), 2.64 (br. s., 1 H), 2.40- 2.06 (m, 7 H), 2.01-1.71 (m, 4H), 1.38-1.25 (m, 1 H), 1.13 (s, 4 H) 385.3 [M + H]⁺ 20

1H NMR (400 MHz, DMSO-d6) δ 10.06-9.79 (m, 2 H), 9.60 (br. s., 1 H),7.45-7.30 (m, 2 H), 7.26-7.15 (m, 3 H), 6.19 (s, 1 H), 5.55 (br. s., 1H), 4.99 (br. s., 1H), 3.95 (br. s., 1 H), 3.84 (d, J = 5.5 Hz, 2 H),3.38- 3.24 (m, 2 H), 3.22-3.10 (m, 1 H), 3.05-2.77 (m, 2 H), 2.65 (br.s., 1 H), 2.33 (br. s., 1 H), 2.29-2.19 (m, 2H), 2.19-2.06 (m, 4 H),2.01- 1.76 (m, 4 H), 1.38-1.24 (m, 1 H), 1.19-1.04 (m, 4 H) 385.3 [M +H]⁺

Compound 21:2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamide

Step 1:

2-(((6-(1R,2S)-2-(E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamide

To a sealed vial charged with2,2,2-trifluoro-N-(2-(2-hydroxy-2-methylpropyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(0.1610 g, 326 μmol) N,N-dimethylformamide, and sodium carbonate (100mg, 943 mol) was added ethenesulfonamide (80.1 mg, 748 μmol) at roomtemperature. The vial was then sealed and heated to 60° C. in an oilbath overnight. The crude mixture was partitioned between2-methyltetrahydrofuran and water. The organic phase was washed twicewith water/brine (1:1 v/v), dried over sodium sulfate and evaporatedunder vacuum to afford crude2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamideThe crude product was used without further purification. LCMS (ESI) m/z:485.9 [M+H]^(+.)

Step 2:

Compound 32:2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamidebis(2,2,2-trifluoroacetate)

Crude2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamidewas dissolved in methanol (2.0 mL) and treated with 10% NaOH aqueous(0.5 mL) at 0° C. for 30 min. The reaction mixture was purified byreverse phase column chromatography 10-30% MeCN/0.1% aqueous TFA aseluent. The pure fractions were combined, frozen and lyophilized overthree days. The desired product2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanesulfonamidebis(2,2,2 trifluoroacetate) was obtained as a white amorphous solid(49.6 mg). LCMS (ESI) m/z: 390.2 [M+H]⁺. ¹H NMR (500 MHz, MeOD) δ7.34-7.28 (m, 2H), 7.23-7.15 (m, 3H), 6.26 (s, 1H), 4.36 (s, 2H), 4.27(s, 2H), 3.92 (p, J=8.1 Hz, 1H), 3.67 (t, J=6.7 Hz, 2H), 3.41 (t, J=6.7Hz, 2H), 2.88-2.79 (m, 2H), 2.79-2.72 (m, 1H), 2.67-2.56 (m, 2H),2.40-2.25 (m, 2H), 2.09-2.01 (m, 1H), 1.40 (s, 2H), 1.33-1.20 (m, 3H),1.17 (t, J=7.6 Hz, 3H).

Using the appropriate starting material and modifications, the followingexamples in Table 4 were synthesized using the synthetic proceduresdescribed for Compound 21. Only the 1R,2S isomers are shown in the tablein an effort to reduce page length.

TABLE 4 LCMS Compound Structure/name 1H NMR m/z 22

1H NMR (500 MHz, MeOD) δ 7.35- 7.28 (m, 2H), 7.24-7.16 (m, 3H), 6.26 (s,1H), 4.45-4.12 (m, 4H), 3.97-3.87 (m, 1H), 3.44 (t, J = 6.4 Hz, 2H),2.92-2.73 (m, 3H), 2.70 (s, 1H), 2.64-2.53 (m, 4H), 2.39-2.25 (m, 2H),2.09-1.99 (m, 1H), 1.32-1.20 (m, 2H), 1.17 (t, J = 7.6 Hz, 3H). 354.4[M + H]⁺ 23

(400 MHz, DMSO-d₆) δ 10.73 (br s, 1H), 9.91-9.73 (m, 2H), 7.38- 7.32 (m,2H), 7.25 (br s, 2H), 7.23- 7.18 (m, 3H), 6.20 (s, 1H), 3.91- 3.82 (m,1H), 3.62-3.57 (m, 2H), 3.46-3.37 (m, 4H), 3.07-2.98 (m, 1H), 2.94-2.86(m, 1H), 2.69- 2.62 (m, 1H), 2.39-2.30 (m, 1H), 2.29-2.21 (m, 3H),2.17-2.08 (m, 3H), 1.99-1.79 (m, 4H), 1.34- 1.26 (m, 1H), 1.14 (app-t, J= 7.4 Hz, 4H) 418.2 [M + H]⁺ 24

1H NMR (400 MHz, DMSO-d6) δ 10.33-10.17 (m, 1 H), 9.90 (br. s., 2 H),7.61 (br. s., 1 H), 7.39-7.28 (m, 2 H), 7.28-7.14 (m, 3 H), 7.09 (br.s., 1 H), 6.19 (s, 1 H), 3.84 (br. s., 1 H), 3.34-3.16 (m, 4 H), 2.92(d, J = 12.3 Hz, 2 H), 2.67-2.55 (m, 3 H), 2.33 (br. s., 1 H), 2.24 (q,J = 7.5 Hz, 2 H), 2.19-2.06 (m, 2 H), 2.00- 1.73 (m, 3 H), 1.37-1.27 (m,1 H), 1.13 (dt, J = 1.3, 7.5 Hz, 4 H) 382.3 [M + H]⁺

Compound 25:N-ethyl-2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

Step 1:

2-(6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptan-2-yl)aceticacid

A solution of2,2,2-trifluoro-N-(2-(2-hydroxy-2-methylpropyl)-2-azaspiro[3.3]heptan-6-yl)-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(800 mg, 1.62 mmol), 2-oxoacetic acid hydrate (298 mg, 3.24 mmol) and 5drops of AcOH in MeOH (16.2 mL) was treated with sodium cyanoborohydride(305 mg, 4.86 mmol). The reaction mixture was stirred at roomtemperature for 1 h. After 1 h, additional 2-oxoacetic acid hydrate wasadded (298 mg, 3.24 mmol) and the reaction was stirred at 45° C. for 1.5h. The mixture was cooled to room temperature and concentrated underreduced pressure. The residue was diluted with DCM then quenched withsat. NaHCO3. The layers were separated and the aqueous was extractedonce with DCM. The combined organic layer was washed once with brine,dried with sodium sulfate and evaporated under reduced pressure toafford crude2-(6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptan-2-yl)aceticacid (522 mg). LCMS m/z 437.2 [M+H]⁺.

Step 2:

N-(2-(2-(ethylamino)-2-oxoethyl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide

To a solution of2-(6-(2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2-azaspiro[3.3]heptan-2-yl)aceticacid (175 mg, 400 μmol), ethylammonium chloride (40.4 mg, 496 μmol) anddiisopropylethylamine (215 μL, 1.24 mmol) in DMF (2.8 mL) at 0° C. wasadded HATU (236 mg, 621 μmol). The reaction mixture was allowed to warmand stirred at room temperature for 3 h. The reaction mixture wasdiluted with water and extracted with ethyl acetate (2×15 mL). Thecombined organic phase was washed once with brine, dried over anhydrousNa₂SO₄ then concentrated to afford crudeN-(2-(2-(ethylamino)-2-oxoethyl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(199 mg, 430 μmol) as a yellow oil with residual DMF. LCMS m/z 464.2[M+H]⁺.

Step 3:

N-ethyl-2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

To a solution ofN-(2-(2-(ethylamino)-2-oxoethyl)-2-azaspiro[3.3]heptan-6-yl)-2,2,2-trifluoro-N-((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)acetamide(190 mg, 409 μmol) in MeOH (2.0 mL) at room temperature was added NaOH(1M, 2.04 mL, 2.04 mmol). The mixture was allowed stirred at roomtemperature for 30 min. The mixture was concentrated under reducedpressure and the crude product was dissolved in TFA/water and purifiedon a 12 g C-18 column using MeCN/0.1% TFA H2O as eluents with thefollowing gradient (2 CV 0%, 20 CV 0-30%, 5 CV 30%) to affordN-ethyl-2-(6-(((1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate) (86.7 mg) as a white solid afterlyophilisation. LCMS m/z 368.1 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 10.23(br s, 1H), 9.23 (br s, 2H), 8.45-8.33 (m, 1H), 7.35 (t, J=7.5 Hz, 2H),7.27-7.14 (m, 3H), 6.19 (s, 1H), 4.27-4.10 (m, 1H), 4.10-3.99 (m, 2H),3.98-3.87 (m, 2H), 3.80-3.63 (m, 1H), 3.16-3.04 (m, 2H), 2.77-2.57 (m,2H), 2.57-2.52 (m, 2H), 2.48-2.35 (m, 2H), 2.30-2.17 (m, 2H), 2.01-1.85(m, 1H), 1.20-1.14 (m, 2H), 1.12 (t, J=7.5 Hz, 3H), 1.03 (t, J=7.2 Hz,3H).

Using the appropriate starting material and modifications the followingexamples in Table 5 were synthesized using the synthetic proceduresdescribed for Compound 25. Only the 1R,2S isomers are shown in the tablein an effort to reduce page length.

TABLE 5 LCMS Compound Structure/name 1H NMR m/z 26

¹H NMR (400 MHz, DMSO-d6) δ 10.26 (br s, 1H), 9.15 (br s, 2H), 8.70 (t,J = 5.7 Hz, 1H), 7.35 (t, J = 7.5 Hz, 2H), 7.27-7.17 (m, 3H), 6.21 (s,1H), 4.31-4.19 (m, 1H), 4.17-4.12 (m, 1H), 4.12-4.05 (m, 2H), 4.04-3.96(m, 1H), 3.52 (q, J = 6.5 Hz, 2H), 3.27 (t, J = 6.6 Hz, 2H), 3.02 (s,3H), 2.75 (s, 1H), 2.71-2.64 (m, 1H), 2.63-2.52 (m, 1H), 2.46- 2.34 (m,2H), 2.29-2.18 (m, 2H), 2.01-1.90 (m, 1H), 1.20- 1.14 (m, 2H), 1.12 (t,J = 7.5 Hz, 3H). 446.3 [M + H]⁺ 27

¹H NMR (400 MHz, DMSO-d6) δ 10.21 (br s, 1H), 9.25 (br s, 2H), 8.31 (d,J = 7.5 Hz, 1H), 7.38-7.32 (m, 2H), 7.26-7.18 (m, 3H), 6.21 (s, 1H),4.29-4.19 (m, 1H), 4.15-4.11 (m, 1H), 4.11-4.04 (m, 2H), 3.96-3.90 (m,2H), 3.84 (sept, J = 6.6 Hz, 1H), 3.80-3.70 (m, 1H), 2.79- 2.65 (m, 2H),2.62-2.52 (m, 1H), 2.47-2.35 (m, 1H), 2.28- 2.20 (m, 2H), 2.02-1.91 (m,1H), 1.22-1.14 (m, 2H), 1.12 (t, J = 5.9 Hz, 3H), 1.07 (d, J = 6.6 Hz,6H). 382.2 [M + H]⁺ 28

¹H NMR (400 MHz, DMSO-d6) δ 10.08 (br s, 1H), 9.16 (br s, 2H), 7.35 (t,J = 7.5 Hz, 2H), 7.27- 7.18 (m, 3H), 6.21 (s, 1H), 4.32- 4.27 (m, 2H),4.27-4.23 (m, 1H), 4.18-4.10 (m, 2H), 4.10- 4.03 (m, 1H), 3.77 (br s,1H), 2.87 (d, J = 8.9 Hz, 6H), 2.79- 2.71 (m, 1H), 2.71-2.64 (m, 1H),2.65-2.54 (m, 1H), 2.48- 2.35 (m, 1H), 2.29-2.18 (m, 2H), 2.01-1.90 (m,1H), 1.20- 1.15 (m, 2H), 1.12 (t, J = 7.6 Hz, 3H). 368.1 [M + H]⁺ 29

¹H NMR (400 MHz, DMSO-d6) δ 10.32 (br s, 1H), 9.18 (br s, 2H), 7.35 (t,J = 7.5 Hz, 2H), 7.29- 7.14 (m, 3H), 6.21 (s, 1H), 4.22 (s, 2H), 4.13(s, 2H), 4.11-4.04 (m, 1H), 3.84-3.71 (m, 1H), 3.18 (d, J = 1.0 Hz, 3H),2.80- 2.70 (m, 1H), 2.70-2.57 (m, 2H), 2.49-2.39 (m, 2H), 2.29- 2.17 (m,2H), 2.02-1.86 (m, 1H), 1.22-1.14 (m, 2H), 1.12 (t, J = 7.5 Hz, 3H).418.3 [M + H]⁺ 30

¹H NMR (400 MHz, DMSO-d6) δ 10.33 (br s, 1H), 9.28 (br s, 2H), 7.95 (s,1H), 7.64 (s, 1H), 7.35 (t, J = 7.5 Hz, 2H), 7.29- 7.16 (m, 3H), 6.21(s, 1H), 4.28- 4.13 (m, 1H), 4.13-3.88 (m, J = 16.5, 9.8 Hz, 3H),3.87-3.70 (m, 1H), 2.81-2.63 (m, J = 20.1 Hz, 2H), 2.62-2.52 (m, 1H),2.46- 2.34 (m, J = 8.4 Hz, 1H), 2.29- 2.18 (m, 2H), 2.04-1.90 (m, 1H),1.27 (d, J = 6.8 Hz, 3H), 1.22-1.14 (m, J = 12.7,5.5 Hz, 2H), 1.12 (t, J= 7.5 Hz, 3H). 354.2 [M + H]⁺ 31

¹H NMR (400 MHz, dmso) δ 9.39 (br s, 1H), 9.18 (br s, 2H), 7.47 (s, 1H),7.39-7.32 (m, 2H), 7.30 (s, 1H), 7.27-7.17 (m, 3H), 6.20 (s, 1H),4.36-4.27 (m, 1H), 4.24-4.08 (m, 3H), 3.82-3.69 (m, 1H), 3.29-3.18 (m,2H), 2.82-2.64 (m, 2H), 2.64-2.53 (m, 1H), 2.50-2.31 (m, 2H), 2.31-2.17(m, 2H), 2.06-1.84 (m, 1H), 1.21-1.15 (m, 2H), 1.14 (s, 6H), 1.11 (t, J= 7.6 Hz, 3H). 382.4 [M + H]⁺

Each of the aforementioned examples can be reproduced to afford theopposite enantiomer (i.e., 1S,2R) using the procedures described aboveand intermediate(1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate instead of(1R,2S)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropan-1-amine(S)-2-hydroxy-2-phenylacetate. The structures of these enantiomers arerepresented in the following table below.

Structure/name

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethanol bis(2,2,2-trifluoroacetate)

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3- diol bis(2,2,2-trifluoroacetate)

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3- diol

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propane-1,3- diol 2-hydroxypropane-1,2,3-tricarboxylate

2-(2-aminoethyl)-N-((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-2- azaspiro[3.3]heptan-6-aminebis(2,2,2-trifluoroacetate)

(R)-1-(2-(((1S,2R)-2-((E)-1-phenylbut- 1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7-yl)propan-2-ol dohydrochloride

7-(2-methoxyethyl)-N-(((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)-7- azaspiro[3.5]nonan-2-aminedihydrochloride

2-methyl-1-(2-(((1S,2R)-2-((E)-1- phenylbut-1-en-2-yl)cyclopropyl)amino)-7- azaspiro[3.5]nonan-7-yl)propan-2-oldihydrochloride

(R)-3-(2-(((1S,2R)-2-((E)-1-phenylbut- 1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7-yl)propane-1,2- diol dihydrochloride

(S)-3-(2-(((1S,2R)-2-((E)-1-phenylbut- 1-en-2-yl)cyclopropyl)amino)-7-azaspiro[3.3]nonan-7-yl)propane-1,2- diol dihydrochloride

N-2-(6-(((1S,2R)-2-((E)-1-phenylbut-1- en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2- yl)ethyl)merthanesulfonamidebis(2,2,2-trifluoroacetate)

N-(2-(6-(((1S,2R)-2-((E)-1-phenylbut-1- en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)acetamide bis(2,2,2-trifluoroacetate)

N-(2-(6-(((1S,2R)-2-((E)-1-phenylbut-1- en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2- yl)ethyl)cyclopropanesulfonamidebis(2,2,2-trifluoroacetate)

N-2-(6-(((1S,2R)-2-((E)-1-phenylbut-1- en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2- yl)ethyl)ethanesulfonamide bis(2,2,2-trifluoroacetate)

N-2-(6-(((1S,2R)-2-((E)-1-phenylbut-1- en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)ethyl)propane- 2-sulfonamidebis(2,2,2-trifluoroacetate)

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2- yl)ethanesulfonamide bis(2,2,2- trifluoroacetate)

3-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propanamide bis(2,2,2-trifluoroacetate)

2-(2-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7- yl)ethanesulfonamide dihydrochloride

3-(2-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7-yl)propanamide dihydrochloride

2-(6-(2,2,2-trifluoro-N-((1S,2R)-2-((E)- 1-phenylbut-1-en-2-yl)cyclopropyl)acetamido)-2- azaspiro[3.3]heptan-2-yl)acetic acid

2-methoxy- N-2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)ethyl)ethanesulfonamide bis(2,2,2-trifluoroacetate)

2-(2-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7-yl)propane-1,3-diol bis(2,2,2-trifluoroacetate)

2-(2-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-7-azaspiro[3.5]nonan-7-yl)ethanol dihydrochloride

2-methyl-1-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propan-2-ol bis(2,2,2-trifluoroacetate)

N-ethyl-2-(6-(((1S,2R)-2-((E)-1- phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

N-(2-(methylsulfonyl)ethyl)-2-(6- (((1R,2S or 1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

N-isopropyl-2-(6-(((1S,2R)-2-((E)-1- phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

N,N-dimethyl-2-(6-(((1S,2R)-2-((E)-1- phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

N-(2-hydroxyethyl)-2-(6-(((1S,2R)-2- ((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

2-methyl-2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propanamide bis(2,2,2-trifluoroacetate)

N-(2-methyl-1-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)propan-2- yl)methanesulfonamide bis(2,2,2-trifluoroacetate)

N-(methanesulfonyl)-2-(6-(((1S,2R)-2- ((E)-1-phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)acetamidebis(2,2,2-trifluoroacetate)

2-(6-(((1S,2R)-2-((E)-1-phenylbut-1-en- 2-yl)cyclopropyl)amino)-2-azaspiro[3.3]heptan-2-yl)proanamide bis(2,2,2-trifluoroacetate)

2,2-dimethyl-3-(6-(((1S,2R)-2-((E)-1- phenylbut-1-en-2-yl)cyclopropyl)amino)-2- azaspiro[3.3]heptan-2-yl)propanamidebis(2,2,2-trifluoroacetate)

LSD1 TR-FRET Assay

LSD1 demethylase reactions were carried out in 50 mM HEPES pH 7.4, 100mM NaCl, 1 mM DTT, 0.01% Tween-20, and 0.1 mg/mL BSA. All enzymaticreactions were performed for 50 minutes at room temperature in a 10-μLvolume. Five microliters of 8 μM biotinylated H3K4me1 peptide solutionwas added to each well of a black 384 well, clear-bottom assay platecontaining 80 nL compound (final concentration of 0.8% DMSO and 4 μMsubstrate). Reactions were initiated by the addition of a mixturecontaining 20 nM LSD1 and 80 nM FAD (5 μL). LSD1 and FAD finalconcentrations were 10 and 40 nM, respectively. Enzyme activity wasstopped by the addition of 90 μL of high salt buffer consisting of 50 mMHEPES pH 7.4, 500 mM NaCl, 1 mM DTT, 0.01% Tween-20, and 0.1 mg/mL BSA.Ten microliters of the quenched reaction mixtures were transferred to ablack 384 well ProxiPlate. Ten microliters of detection mixture wasadded to the ProxiPlate, Europium-labeled antibody and Streptavidin APCwere used at final concentrations of 0.3 nM and 200 nM, respectively(total assay volume of 20 μL). Capture of the product peptide by theanti-H3K4me0 antibody and Streptavidin APC was allowed to proceed for 60min at room temperature before measuring the TR-FRET signal. Plates wereread on a Perkin Elmer EnVision. Percent inhibition was calculated usingMax (no inhibitor) and Min (quenched with stop buffer) controls andinhibition curves plotted to determine IC₅₀ values.

LSD1 LY96 Quantigene Assay

MV4-11 cells were cultured at a density of 4×104 cells per well in a96-well plates and treated with various doses inhibitor starting from 10μM up to 0.0005 μM for 16 h. The LY-96 mRNA induction was quantifiedusing the Quantigene 2.0 system (Affymetrix). The cells were lysed withLysis Mixture containing Proteinase K. The working reagent for capturingthe RNA was prepared according to the steps detailed in “CapturingTarget RNA from Cultured Cell or Blood Lysates” in the Quantigenehandbook. The subsequent hybridization with LY-96 probe, signalamplification and detection steps were performed as described in themanual. The chemiluminescence was read using Envision (PerkinElmer) andAbase (IDBS software) was used to plot the dose response curves andcalculate IC₅₀.

Kasumi-1 G1₅₀ Assay

Cells were plated at 5,000 cells per well in 96 well tissue culturedishes containing tool compounds arrayed in a 10-point dose curve,ranging from 0 to 10 mM with 4-fold dilutions, and split every fourthday at a fixed ratio to re-establish 5,000 cells/well density forDMSO-treated controls. Cell treatments were carried out for a total of12 days. At each 4-day split, the viable cell numbers were determinedusing the Cell Titer-Glo luminescent cell viability assay (Promega,Madison, Wis. USA) using an EnVision® Multilabel Plate Reader (PerkinElmer, Waltham, Mass. USA). GraphPad Prism 6 (GraphPad Software, Inc.,La Jolla, Calif. USA) was used for curve fitting and determination ofGI₅₀ values.

TABLE 6 Com- LSD1 TR-FRET LSD1 LY96 Kasumi-1 pound IC₅₀ EC₅₀ GI₅₀  1 A AA  2 A A A  4 A A A  5 ND A A  6 ND A A  7 ND A A  8 ND A A  9 ND A A 10ND A A 11 A A A 13 A A A 14 ND A A 15 ND A A 16 ND A A 17 A A A 18 ND AA 19 ND A A 20 ND A A 22 A A A 23 A A A 24 ND A A 26 ND A A 27 ND A A 28ND A A 29 ND A A 30 ND A A 32 A A A *IC₅₀, EC₅₀ and GI₅₀ values arereported as follows: “A” indicates an IC₅₀ value of less than 100 nM;“B” indicates an IC₅₀ value of 100 nM to 1 μM; “C” indicates an IC₅₀value of greater than 1 μM and less than 10 μM for each enzyme; “D”indicates an IC₅₀ value of greater than 10 μM for each enzyme; “*(X μM)”indicates that no inhibition was observed at the highest concentration(i.e., X μM) of compound tested; and “ND” is not determined.

The invention claimed is:
 1. A compound of the Formula:

or a pharmaceutically acceptable salt thereof.
 2. The compound of claim1, wherein the compound is of the Formula:

or a pharmaceutically acceptable salt thereof.
 3. The compound of claim2, wherein the enantiomeric purity of the compound is at least 97%. 4.The compound of claim 2, wherein the enantiomeric purity of the compoundis at least 98%.
 5. The compound of claim 2, wherein the enantiomericpurity of the compound is at least 99%.
 6. A pharmaceutical compositioncomprising an effective amount of a compound of claim 1, or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 7. A pharmaceutical composition comprising aneffective amount of a compound of claim 2, or a pharmaceuticallyacceptable salt thereof; and a pharmaceutically acceptable carrier.
 8. Amethod of treating a disease that is treatable by inhibiting LSD1wherein said disease is selected from HIV, prostate cancer, esophagealsquamous cell, neuroblastoma, ovarian cancer, bladder cancer, lungcancer, colorectal cancer, squamous cell carcinomas, breast cancer,glioblastoma multiforme, chondrosarcoma, Ewing's sarcoma, osteogenicsarcoma, rhabdomyosarcoma, melanoma, or medulloblastoma comprising thestep of administering to a subject in need thereof a compound of claim1, or a pharmaceutically acceptable salt thereof.
 9. A method oftreating a disease that is treatable by inhibiting LSD1 wherein saiddisease is selected from HIV, prostate cancer, esophageal squamous cell,neuroblastoma, ovarian cancer, bladder cancer, lung cancer, colorectalcancer, squamous cell carcinomas, breast cancer, glioblastomamultiforme, chondrosarcoma, Ewing's sarcoma, osteogenic sarcoma,rhabdomyosarcoma, melanoma, or medulloblastoma comprising the step ofadministering to a subject in need thereof a compound of claim 2, or apharmaceutically acceptable salt thereof.
 10. A compound having theFormula:

or a pharmaceutically acceptable salt thereof.
 11. The compound of claim10, wherein the compound is of the Formula:

or a pharmaceutically acceptable salt thereof.
 12. The compound of claim11, wherein the enantiomeric purity of the compound is at least 97%. 13.The compound of claim 11, wherein the enantiomeric purity of thecompound is at least 98%.
 14. The compound of claim 11, wherein theenantiomeric purity of the compound is at least 99%.
 15. Apharmaceutical composition comprising an effective amount of a compoundof claim 10, or a pharmaceutically acceptable salt thereof; and apharmaceutically acceptable carrier.
 16. A pharmaceutical compositioncomprising an effective amount of a compound of claim 11, or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 17. A method of treating a disease that is treatableby inhibiting LSD1 wherein said disease is selected from HIV, prostatecancer, esophageal squamous cell, neuroblastoma, ovarian cancer, bladdercancer, lung cancer, colorectal cancer, squamous cell carcinomas, breastcancer, glioblastoma multiforme, chondrosarcoma, Ewing's sarcoma,osteogenic sarcoma, rhabdomyosarcoma, melanoma, or medulloblastomacomprising the step of administering to a subject in need thereof acompound of claim 10, or a pharmaceutically acceptable salt thereof. 18.A method of treating a disease that is treatable by inhibiting LSD1wherein said disease is selected from HIV, prostate cancer, esophagealsquamous cell, neuroblastoma, ovarian cancer, bladder cancer, lungcancer, colorectal cancer, squamous cell carcinomas, breast cancer,glioblastoma multiforme, chondrosarcoma, Ewing's sarcoma, osteogenicsarcoma, rhabdomyosarcoma, melanoma, or medulloblastoma comprising thestep of administering to a subject in need thereof a compound of claim11, or a pharmaceutically acceptable salt thereof.